Composition and method

ABSTRACT

A process, and the composition used therein, for estrogenic therapy which comprises administering daily to the patient from about 0.2 to 2000 milligrams of a compound having the formula: ##EQU1## wherein R is selected from the group consisting of hydrogen, lower alkyl and a lower saturated acyclic acyl; Z is selected from the group consisting of &gt;CH 2 , &gt;CHOH and &gt;C=O; and A is selected from the group consisting of --CH 2  --CH 2  -- and --CH=CH--.

This is a continuation of application Ser. No. 441,150, filed Feb. 11, 1974; which in turn is a continuation of application Ser. No. 289,456, filed Sept. 15, 1972; which in turn is a continuation of application Ser. No. 28,913, filed Apr. 15, 1970; which in turn is a continuation-in-part of application Ser. No. 512,199, filed Dec. 7, 1965 all of which are abandoned.

The present invention is directed to a pharmaceutical composition useful in carrying out estrogenic therapy for human patients.

The administration of estrogenic hormones such as estradiol and estrogenic substances such as diethylstilbestrol in the therapeutic treatment of various illnesses and disorders such as acne, capillary fragility, emotional instability, delayed puberty, reduction of blood cholesterol and habitual abortion, has been practiced for years with considerable success. The estrogen hormones and estrogenic substances currently available are not without criticism, however. A major drawback of these substances, particularly as applied to their administration in men, is their strong "feminizing" effect. Many women also manifest undesirable side-effects from these compounds, as, for example, excessive menstrual flow and masculinization. A search has been in progress for years with little success for a "weak estrogen" compound which is effective in estrogenic therapy but lacks distinct feminizing and other undesirable side effects.

In accordance with the present invention, there is provided a pharmaceutical composition containing an effective therapeutically active estrogenic ingredient characterized by relatively weak feminizing and other side effect estrogenic activity.

The active ingredient of the pharmaceutical compositions can be represented by the structural formula: ##EQU2## wherein R is hydrogen, substituted or unsubstituted alkyl, e.g. lower alkyl such as methyl, ethyl, hexyl, etc., and acyl, e.g., lower saturated acyclic acyl radicals such as acetyl and valeryl; A is --CH₂ --CH₂ -- or --CH=CH-- and Z is >C=O, >CH₂ or >CHOH. As will be demonstrated below, the compounds of the invention when compared with diethylstilbestrol, a compound of known estrogenic activity used in estrogenic therapy in the mouse uterotropic activity test, exhibit effective but relatively weak estrogenic activity.

The compounds of the present invention include the compound: ##EQU3## hereinafter referred to as the fermentation estrogenic substance (F.E.S.), from which the other compounds of the invention can be produced by reduction of the ketone group to replace the oxygen of the ketone group with two hydrogen atoms, by reducing the ketonic group to add two hydrogens thereto, by saturation of the olefinic bond or any combination of such reductions. The reduction of the ketone group to replace the oxygen can be effected by several procedures. One of these procedures involves the Clemmensen reduction using zinc and hydrochloric acid; another involves the Wolff-Kishner reduction using hydrazine and alkali, e.g. NaOH, and the third involves formation of the dithioacetal with ethylene dithiol or ethylmercaptan and the catalytic desulfurization with Raney nickel catalyst containing adsorbed hydrogen.

The addition of two hydrogen atoms to the ketonic group and saturation of the olefinic bond can be obtained by conventional reduction procedures, for instance, in the presence of Raney nickel catalyst. The reduction is preferably carried out with the F.E.S. suspended or dissolved in a suitable solvent, e.g. an alcohol, preferably a lower alkanol such as methanol, ethanol, etc. In general, the reduction can be accomplished at ambient temperatures and ambient pressures. Preferable temperatures are from about 15° to 40°C., and preferable pressures are of from about 1 to 100 atmospheres. In general, from about 0.1 to 5 grams of catalyst are used per gram of F.E.S.

In producing compounds of the invention where A is --CH₂ --CH₂ -- the olefinic bond of F.E.S. can be reduced, for example, by hydrogenation in the presence of a Group VIII metal, particularly platinum or palladium catalyst on a suitable carrier, e.g., charcoal. Generally, the catalyst contains from about 0.01 to about 10% of the catalytic metal. The catalyst is used in a ratio of generally from 0.02 to 2 grams of catalyst, preferably about 0.1 to 0.5 gram, and particularly about 0.2 gram catalyst per gram of F.E.S. The reduction may be carried out while the F.E.S. is dissolved in a suitable solvent, e.g. an alcohol, especially a lower alkanol such as 2-propanol, methanol, ethanol, and acid, e.g. acetic acid, etc. at ambient temperatures; e.g. from about 15° to 40° C., and ambient pressures, since only the presence of hydrogen is required; however, it is preferred to utilize an elevated pressure, e.g. from about 1 to 50 atmospheres of hydrogen.

In producing compounds of the present invention where R is alkyl, conventional alkylation procedures can be used to replace the H atom of one or both of the OH groups on the benzene ring of F.E.S. with an alkyl group. Alkylated dihydro F.E.S. compounds can be produced, for example, by first alkylating F.E.S. and then reducing it as set forth supra, or by first reducing it and then alkylating it. The alkylation can be by reaction with the corresponding dialkyl sulfates, e.g. dimethyl sulfate, diethyl sulfate, etc., to produce the dialkyl F.E.S. or a monoalkyl F.E.S. with the alkyl group replacing the hydrogen of the hydroxyl group on the benzene ring ortho to the ester group. Furthermore, a monomethyl F.E.S. compound with the methyl group replacing the hydrogen of the hydroxyl group para to the ester group can be selectively produced using diazomethane.

In producing compounds of the present invention where R is acyl, conventional acylation procedures can be used to replace the hydrogen atom of one or both of the hydroxyl radicals on the benzene ring of F.E.S. with an acyl radical. Acylated F.E.S. compounds can be produced, for example, by reaction with the corresponding acid anhydride, e.g. acetic anhydride, propionic anhydride, etc., catalyzed with, for example, sodium acetate or pyridine. Ambient conditions can be used although it is preferred to keep the reaction mixture cold. When compounds having one R as alkyl and the other acyl, it is advantageous to alkylate before acylating.

The fermentation estrogenic substance (F.E.S.) is so named since a convenient method for producing it is by cultivating, on a suitable nutrient medium, the organism Gibberella zeae (Gordon) on deposit at the Northern Utilization Research and Development Division of the United States Department of Agriculture under the number NRRL-2830.

Specific examples of the preparation of F.E.S. and other compounds of the invention are given below and disclosed in more detail in U.S. Pat. Nos. 3,196,019; 3,239,354; 3,239,345 and 3,239,341.

The pharmaceutical compositions of the invention can be prepared by mixing the active ingredient with non-toxic, pharmaceutically-acceptable carriers, which can be inert diluents or solid carriers, and forming the resulting mixture into suitable dosage unit forms. The compositions can be administered to the subject by any suitable method including oral and parenteral administration. Forms suitable for oral administration include, for example, pressed or coated tablets, capsules or pills, syrups, solutions or suspensions in water or non-toxic organic solvent media such as propylene glycol and glycerol formal, and dispersible powders. Compositions suitable for parenteral administration are the known pharmaceutical forms for such administrations, for example, sterile aqueous suspensions or solutions in oily media. The sterile aqueous suspensions can be formulated in the presence of parenterally acceptable buffers, e.g. sodium citrate, citric acid and/or preservatives such as phenol and methyl and propyl esters of p-hydroxy benzoic acid. A preferred oily media for preparation of the sterile aqueous solution is peanut oil. For treatment of the skin, the active ingredient can be mixed in any of the conventional cosmetic base lotions in a hydrophilic base.

The pharmaceutical compositions may also include adjuvants known in the art as desirable or useful as, for example, wetting agents, dispersing agents, suspending agents, lubricating agents, sweetening agents, coloring agents and flavoring agents.

Illustrative of oral compositions are tablets wherein the active ingredient is mixed with inert fillers, e.g., dicalcium phosphate, terra alba or lactose in the presence of disintegrating agents as, for example, maize starch and in the presence of lubricating agents such as magnesium stearate. Examples of suitable aqueous solutions for oral use are those formulated by incorporating the active ingredient in inert pharmaceutically-acceptable liquid solvent media which can contain, if desired, pharmaceutically-acceptabe thickening agents such as sodium carboxy-methyl-cellulose and/or pharmaceutically-acceptable sweetening and flavoring agents.

The actual amounts of the active ingredient in the pharmaceutical composition of the invention may vary depending on the particular disorder treated but in all cases the amount present is that sufficient to produce the desired therapeutic effect. In general, for carrying out estrogenic therapy for a human patient from about 0.2 to 2000 milligrams of the active ingredient are administered daily. This amount is administered in amounts of from about 0.2 to 500 mg. preferably 1.0 to 100 mg. per dosage unit. More particularly, in post-menopause usage the active ingredient will be administered in a range of about 0.1 to 10 mg., preferably about 0.5 or 1 to 6 mg. per day per kilogram body weight of the patient. With females this dosage is typically administered in a manner similar to oral contraceptives, that is, 20 to 25 days on the drug followed by rest periods of up to ten days, e.g. 5 to 10 days. Often this cycle is referred to as 3 weeks on the drug and 1 week rest period. Post-menopause usage and the dosages used therefore includes treatment of capillary fragility and emotional instability and such dosages are those used with habitual abortion, delayed puberty, etc. The cycle regime is used to to avoid building of endometrium as is well known. The dosage for treatment of cholesterol, i.e. for cholesterol lowering, is from about 0.5 to 20 mg./day/kilogram of body weight and the dosage is usually given continuously although cyclic administration can be used. Acne is treated with a skin cream containing the active ingredient, e.g. 0.1 to 10 wt. % of the active ingredient in a cosmetic base lotion or in a hydrophilic base.

The following examples are offered to illustrate this invention; however, the invention is not limited to the specific materials, amounts and procedures set forth. The first example illustrates preparation of a suitable inoculum containing the organism Gibberella zeae (Gordon) NRRL-2830.

EXAMPLE I

A spore sand culture containing Gibberella zeae (Gordon) NRRL-2830 was aseptically placed in a sterile tube containing 15 milliliters of Czapek's-Dox solution and a small amount of agar. This medium was then incubated for about 168 hours at approximately 25°C. At the end of the incubation period, the medium was washed with 5 milliliters of sterile deionized water and transferred to a sterile tube containing 45 milliliters of Czapek's-Dox solution. The contents of the tube were then incubated for about 96 hours at about 25°C. after which the material was available for use in inoculation of a fermentation medium.

The following example illustrates the fermentation of the organism Gibberella zeae (Gordon) NRRL-2830 to produce F.E.S.

EXAMPLE II

To a 2 liter flask was added 300 grams of finely divided corn. The flask and its contents were then sterilized and after sterilization 150 milliliters of sterile deionized water were added. To the mixture in the flask were then added 45 milliliters of the inoculum prepared by the process of Example I and the material was thoroughly mixed. The mixed material was then incubated for about 20 days at 25°C. in a dark room in a water-saturated atmosphere.

The following example illustrates the recovery of the F.E.S. from the fermentation medium.

EXAMPLE III

A 300 gram portion of fermented material produced by the method of Example II was placed in 500 milliliters of deionized water and slurried. The slurry was then heated for about 15 minutes at 75°C., 300 grams of filter aid were then added and the material was filtered. The solid filtered material containing the F.E.S. was then air dried, and 333 grams of the dried cake were then extracted with 500 milliliters of ethanol. This procedure was repeated three more times. The ethanol extract was evaporated to dryness under vacuum to give 6.84 grams of solid material. This solid material was then dissolved in 20 milliliters of chloroform and extracted with 30 milliliters of an aqueous solution containing 5% by weight of sodium carbonate having an adjusted pH of about 11.2. The extraction process was repeated seven more times. The pH of the sodium-carbonate extract was then adjusted to 6.2 with hydrochloric acid, to yield a F.E.S. substance-containing precipitate. The precipitate and the aqueous sodium-carbonate extract were then each in turn extracted with 75 milliliters of ethyl ether. This procedure was repeated three more times to yield a light yellow ethereal solution, which was then evaporated to yield 116 milligrams of solid F.E.S. This material was then subjected to multiple transfer countercurrent distribution using 100 tubes and a solvent system consisting of two parts chloroform and two parts carbon tetrachloride as the lower phase and four parts methanol and one part water as the upper phase, all parts by volume. The solid material obtained from the multiple transfer countercurrent distribution was F.E.S.

The following examples, Examples IV to VI, illustrate the reduction of F.E.S. to produce tetrahydro F.E.S. having the formula: ##EQU4##

EXAMPLE IV

Tetrahydro F.E.S. was produced by dissolving 0.5 gram F.E.S. in 200 milliliters of ethanol. The F.E.S. was reduced by contacting the solution with hydrogen for 3 hours at 30°C. with 1000 psi using 2 grams of Raney nickel as a catalyst. After filtering and concentrating the reaction mixture, the product was washed with 2 to 3 milliliters of 2-nitropropane are crystallized. It was found to have a melting point from 143°-160°C.

EXAMPLE V

The reduction of F.E.S. was conducted in methanol at 30°C. and 1000 psi hydrogen pressure for 5 hours using Raney nickel catalyst to provide a product melting, after several crystallizations from 2-nitropropane and nitromethane, at 141°-143°C. and analyzing:

    Calc.      (C.sub.18 H.sub.26 O.sub.5)                                                                    Found                                               ______________________________________                                         % C        67.1            67.2                                                % H        8.14            8.28                                                ______________________________________                                    

EXAMPLE VI

The reduction of 1 gram of F.E.S. was conducted in 150 cc. of ethanol at room temperature and 50 psi of hydrogen for 4 hours in the presence of a small amount of Raney nickel (about 1 cc. of a thick suspension in water). The product was concentrated, treated with 5 milliliters of isopropyl alcohol, cooled and filtered. The filtrate was mixed with 5 milliliters of water, left standing over night, cooled and filtered to provide 0.65 gram of product having a melting point of 147°-157°C. This product was recrystallized from isopropyl alcohol-water mixtures two times to provide 0.18 gram of a product having a melting point of 178°-180°C. A product having a melting point of 146°-148°C. and weighing 0.22 gram was also recovered from the filtrate after the first recrystallization of the product weighing 0.65 gram. The reduction of the ketone group introduces an asymmetric carbon atom and makes diastereoisomers possible. The optical activities of the two products were (1 ) for THFES(HM), the product with a melting point of 178°-180°C., [α]_(D) ²⁵ = about + 46° e.g. and (2) for a combination of the THFES(HM) and THFES(LM), the combination product having a melting point of 146°-148°C., [α]_(D) ²⁵ = about +39° e.g. where [α] = α .100/c.1, c = 1% in methanol and 1=2 dm. This product is actually a mixture (about 1:2) of the product melting at 178°-180°C. and its isomer. The low melting isomer can be obtained in pure form by recrystallization out of the combination product using glacial acetic acid to provide pure THFES(LM) which melts at ˜ 155°C. and has optical activity [α]_(D) ²⁵ =+36°.

The following example illustrates the preparation of deoxy tetrahydro F.E.S.

EXAMPLE VII

Two 10 gram portions of F.E.S., each in 200 milliliters acetic acid, were catalytically reduced at room temperature in the presence of 1.2 grams of PdO catalyst at a hydrogen pressure of about 45 psi. The combined reduction mixtures were heated to boiling, filtered, and the filter cake was washed with 50 milliliters of hot acetic acid. The cooled filtrate was added, with stirring, to 2 liters of water. The mixture was stirred for 15 minutes and the white solid was collected by filtration, washed and dried in a vacuum desiccator to yield 19.1 grams of dihydro F.E.S. in which the ethylenic unsaturation is saturated, and having a melting point of 191°-193°C.

The dihydro F.E.S. (1 gram) is added slowly with cooling (ice-bath), to a mixture of 5 cc. of ethylene dithiol 0.25 gram of freshly fused zinc chloride and 2 grams of anhydrous sodium sulfate, contained in a microflask. The mixture is maintained at 5°C. for 20 hours and then at room temperature for 4 hours, whereupon it is poured into 50cc. of ice and the precipitate is collected and subjected to hydrogenolysis. To the reaction product is added 100 cc of 90% ethanol and 15 grams of Raney nickel catalyst and the mixture is refluxed until the reaction is complete. The nickel is removed by centrifugation and is washed several times with hot ethanol by centrifugation followed by decantation, and the centrifugates are combined. The mixture is evaporated to dryness and the residue is suitably recrystallized to yield deoxy THFES having the formula: ##SPC1##

EXAMPLE VIII

Nitrosomethylurea in an amount of 1.2 grams was slowly added to a cold mixture of 3.6 milliliters of 50% potassium hydroxide and 17 milliliters of ether. After a few minutes the yellow ether layer of the mixture was decanted, dried over potassium hydroxide, and then added to a solution of 0.30 grams F.E.S. in 17 milliliters of ether. The resulting yellow mixture was left overnight in a loosely stoppered flask and then ether and diazomethane were evaporated using a steam bath. The remaining gummy residue was crystallized by adding 3 milliliters of water, heating to 60°C., and adding ethanol almost to solution. On cooling, crystals formed yielding 0.137 gram of a product having a melting point of 111°-122°C. and analyzing:

    Calc.      (C.sub.19 H.sub.24 O.sub.5)                                                                    Found                                               ______________________________________                                         %    C         68.7            68.3                                            %    H         7.28            7.38                                            %    OCH.sub.3 9.34            9.17                                            ______________________________________                                    

The p methyl F.E.S. is substituted for the F.E.S. in following essentially the same procedure used in Example VII to produce a compound having the formula: ##SPC2##

The following example illustrates the production of dimethyl F.E.S. and monomethyl F.E.S. derivatives, the monomethyl F.E.S. derivative having the hydrogen in the hydroxyl group ortho to the ester group replaced with a methyl group.

EXAMPLE IX

Dimethyl sulfate (5 milliliters) was added to a solution of 2.24 grams of F.E.S. in 80 milliliters 10% NaOH and 20 milliliters water. The mixture was stirred for one-half hour at 18°-20°C. (cooling bath) and an additional 5 milliliters of dimethyl sulfate was added. After an additional 70 minutes of stirring at 20°-26°C., the solid precipitate, Solid A, was collected by filtration, washed with water and dried in a vacuum desiccator. The filtrate from Solid A was acidified with 25 milliliters 12 N H₂ SO₄ to yield a second precipitate, Solid B, which was collected, washed with water, and dried.

Solid A (0.79 gram having a melting point of 114°-118°C.) was recrystallized from a mixture of 10 milliliters water and 15 milliliters ethanol to yield 0.66 gram of dimethyl F.E.S. having a melting point of 108°-110°C.

Solid B (1.39 grams having a melting point of 152°-162°C.) was recrystallized twice from a mixture of water and alcohol to yield 0.8 gram of monomethyl F.E.S. having a melting point of 169°-174°C. and the following analysis of recrystallized Solid B (monomethyl F.E.S.) was obtained:

    Calc.      (C.sub.19 H.sub.24 O.sub.5)                                                                    Found                                               ______________________________________                                         %    C         68.65           67.97                                           %    H         7.28            7.16                                            %    OMe       9.34            9.28                                            ______________________________________                                    

Each of the o methyl F.E.S. and the dimethyl F.E.S. is substituted for the F.E.S. in the procedure of Example VII to produce the respective compounds: ##SPC3##

and ##SPC4##

The following example illustrates the production of an acylated monomethyl F.E.S. derivative.

EXAMPLE X

To a solution of 368 milligrams of p methyl F.E.S. in 8 milliliters pyridine is added 5 milliliters acetic anhydride and the mixture is held at room temperature for 16 hours. Twenty-five milliliters of water are then added. The mixture is stored in a refigerator for 2 hours. The solid precipitated is collected by filtration, washed with water and dried in a vacuum desiccator to recover a compound which is substituted for the F.E.S. in the procedure of Example VII to produce a compound of the formula: ##SPC5##

which is recovered.

EXAMPLE XI

The compound: ##SPC6##

is produced by substituting o methyl F.E.S. for the dihydro F.E.S. in the procedure of Example VII.

The following example illustrates the reduction of F.E.S. to produce dihydro F.E.S. having the structure: ##EQU5##

EXAMPLE XII

Two 10-gram portions of F.E.S. each in 200 milliliters acetic acid were catalytically reduced at room temperature in the presence of 1.2 grams of PdO catalyst at a hydrogen pressure of about 45 psi. The combined reduction mixtures were heated to boiling, filtered, and the filter cake was washed with 50 milliliters of hot acetic acid. The cooled filtrate was added, with stirring, to 2 liters of water. The mixture was stirred for 15 minutes and the white solid was collected by filtration, washed and dried in a vacuum desiccator to yield 19.1 grams of dihydro F.E.S. having a melting point of 191°-193°C.

The production of dimethyl dihydro F.E.S. is illustrated by the following Example.

EXAMPLE XIII

Dihydro F.E.S. (556 milligrams) was dissolved in 25 milliliters 10% NaOH and 10 milliliters water and the solution was stirred. To the stirred solution was added three, two-milliliter portions of dimethyl sulfate at half-hour intervals followed by stirring for an additional hour. The mixture was acidic and it was made alkaline by the addition of 10 milliliters 10% NaOH and the alkaline mixture was stirred one-half hour. The solid formed was collected by filtration, washed with water and dried in a vacuum desiccator. The product weighed 526 milligrams and melted at 115°-117°C. Recrystallization from a mixture of 10 milliliters of water and 25 milliliters of ethanol provided 371 milligrams of material having a melting point of 124°-125.5°C. It was analyzed with the following results:

    Calc.      (C.sub.20 H.sub.28 O.sub.5)                                                                    Found                                               ______________________________________                                         %    C         68.95           69.02                                           %    H         8.10            8.12                                            %    CH.sub.3 O                                                                               17.81           17.81                                           ______________________________________                                    

The following example illustrates the production of monomethyl and dimethyl dihydro F.E.S., the monomethyl dihydro F.E.S. having a methyl group which replaced the hydrogen of the hydroxyl group on the benzene ring ortho to the ester group.

EXAMPLE XIV

Dimethyl sulfate (5 ml.) was added to a solution of 2.24 g. F.E.S. in 80 ml. of a 10% NaOH solution and 20 ml. of water. The mixture was stirred for one-half hour at 18°-20°C. (cooling bath) and an additional 5 ml. of dimethyl sulfate was added. After an additional 70 minutes of stirring at 20°-26°C., the solid precipitate, Solid A, was collected by filtration, washed with water and dried. The filtrate from Solid A was acidified with 25 ml. 12N H₂ SO₄ to yield a second precipitate, Solid B, which was collected, washed with water, and dried.

Solid A (0.79 g. having a melting point of 114°-118°C.) was recrystallized from a mixture of 10 ml. water and 15 ml. ethanol to yield 0.66 g. of dimethyl F.E.S. having a melting point of 108°-110°C.

Solid B (1.39 g. having a melting point of 152°-162°C.) was recrystallized twice from a mixture of water and alcohol to yield 0.80 g. of monomethyl F.E.S. product having a melting point of 169°-174°C. Analysis of Solid B showed:

    Calc.      (C.sub.19 H.sub.24 O.sub.5)                                                                    Found                                               ______________________________________                                         %    C         68.65           67.97                                           %    H         7.28            7.16                                            %    OMe       9.34            9.28                                            ______________________________________                                    

The olefinic bond of each of the dimethyl F.E.S. and monomethyl F.E.S. is reduced using 50 psi of hydrogen and a small amount of 5% Pd on charcoal catalyst in ethanol and conducting the reduction for 3 hours.

EXAMPLE XV

Monomethyl F.E.S. with the methyl group replacing the hydrogen of the hydroxyl group on the benzene ring para to the ester group was prepared by the following procedure:

Nitrosomethylurea in an amount of 1.2 grams was slowly added to a cold mixture of 3.6 milliliters of 50% potassium hydroxide and 17 milliliters of ether. After a few minutes the yellow ether layer of the mixture was decanted, dried over potassium hydroxide, and then added to a solution of 0.30 grams F.E.S. in 17 milliliters of ether. The resulting yellow mixture was left overnight in a loosely stoppered flask and then ether and diazomethane were evaporated off using a steam bath. The remaining gummy residue was crystallized by adding 3 milliliters of water, heating to 60°C., and adding ethanol almost to solution. On cooling, crystals formed, yielding 0.137 grams of a product having a melting point of 111°-116°C. which was again recrystallized in the same way to yield 0.082 grams of monomethyl F.E.S. having a melting point of 120°-122°C. and the following analysis:

    Calc.      (C.sub.19 H.sub.24 O.sub.5)                                                                    Found                                               ______________________________________                                         %    C         68.7            68.3                                            %    H         7.28            7.38                                            %    OCH.sub.3 9.34            9.17                                            ______________________________________                                    

The olefinic bond of this compound is reduced according to the procedure of Example IX.

The following example demonstrates the relatively low estrogenic activity of the compounds of the pharmaceutical compositions of the invention.

EXAMPLE XVI

A solution of each test compound identified in the table below was admixed with a standard pulverized mouse ration and the solvent was removed by evaporation to provide a dry ration containing the level of test compound per gram of feed indicated ithe table below. A control ration and each test ration were fed to 5 to 10 ovariectomized mice weighing about 20 to 23 grams each at a level of 3 grams per day for a period of 5 days after which the mice were sacrificed and their uteri weighed. An increase in the weight of the uteri from the animal fed the test compounds over the weight of the uteri in the control animals demonstrates estrogenic activity for the test compound.

The control animals showed a percent uterine weight to body weight of about 0.048 to 0.050. A uterine response of 0.060 being just significant, the dose required to give a response of 0.060 then gives the relative uterotrophic activity. The following results were obtained from curves drawn plotting dose/uterus weight as percent body weight.

                         Uterotrophic                                                                   Activity Relative To                                                  Estimated                                                          Compound    M.E.D.*        DES       FES                                       ______________________________________                                         DES+        0.003    μg/g feed                                                                             1       2000                                    HMTHFES.sup.(1)                                                                            1.25               .0024   4.8                                     LMTHFES.sup.(2)                                                                            2.25               .0013   2.7                                     DeoxyTHFES.sup.(3)                                                                         6.25               .00048  0.96                                    Dimethyl FES.sup.(4)                                                                       100.0    (estimated)                                                                              .000003 0.06                                    4-methyl FES.sup.(5)                                                                       100.0    (estimated)                                                                              .000003 0.06                                    2-methyl FES.sup.(6)                                                                       28.0               .000017 0.22                                    Dihydro FES.sup.(7)                                                                        3.0                .0010   2.0                                     FES         6.0                .0005   1.0                                     ______________________________________                                          + diethylstilbestrol                                                            *minimal effective dosage                                                     .sup.(1) high melting tetra hydro FES prepared as in Example VI                .sup.(2) low melting tetra hydro FES prepared as in Example V                  .sup.(3) prepared as in Example VII                                            .sup.(4) prepared as in Example IX                                             .sup.(5) prepared as in Example VIII                                           .sup.(6) prepared as in Example IX                                             .sup.(7) prepared as in Example XII                                      

EXAMPLE XVII

Pharmaceutical preparation containing the compound of Example IV in the form of tablets suitable for administration to human patients;

246 grams of THFES(HM) is triturated with 60 grams of lactose to form an homogeneous powder. To the powder is added 20 grams of silicic acid with hydrolyzed starch and water and the mixture stirred until a homogeneous paste is formed. The paste is then dried and tabletted with 2 grams magnesium stearate to form tablets containing approximately 150 mg. of active ingredient. Similar compositions including F.E.S., THFES(LM), deoxy THFES and the alkylated and acylated compounds can be prepared by substitution of these compounds for THFES(HM) as the active ingredient.

EXAMPLE XVIII

Pharmaceutical preparation of aqueous suspension for oral administration:

    Recipe for 1000 ml. of suspension                                              ______________________________________                                         Compound of Example III                                                                           30.0       g.                                               Sucrose            400.0      g.                                               Powdered tragacanth                                                                               7.5        g.                                               Flavoring essential oil                                                                           0.2        ml.                                              Methyl p-hydroxybenzoate                                                                          2.0        g.                                               Propyl p-hydroxybenzoate                                                                          0.5        g.                                               Glycerol           150.0      ml.                                              Citric acid        2.0        g.                                               Benzoic acid       1.0        g.                                               Distilled water    (to complete 1000 ml.)                                      ______________________________________                                    

The glycerol, benzoic acid, methyl and propyl benzoic acids, tragacanth gum, flavoring oil and active ingredient are mixed into a homogeneous mass. An aqueous solution of the citric acid is then added with slurring and finally the sucrose is added. Slurring is continued until an homogeneous suspension is obtained to which is added the balance of the water. Similar compositions including F.E.S., THFES(LM), deoxy THFES and the alkylated and acylated compounds can be prepared by substitution of these compounds for THFES(HM) as the active ingredient.

EXAMPLE XIX

100 mg. of the compound of Example III, 0.2 mg. of methyl-p-methoxybenzoate, 0.5 mg. of sodium citrate and 0.2 mg. of citric acid are added to 1 ml. of water. The pH of the suspension is adjusted to 5 with HCl. Heat sterilization results in an aqueous suspension suitable for parenteral injection.

EXAMPLES XX - XXIII

Suitable pharmaceutical compositions can be prepared by replacing the F.E.S. in the preparations of Examples XVII-XIX with any one of the compounds of Examples IV-XV.

The following examples demonstrate the cholesterol lowering effect of the active ingredients of this invention.

EXAMPLE XXIV

Male Wistar rats (Royal Hart) were allowed to eat powdered Purina Laboratory Chow ad-lib for 7 days, in order to become accustomed to eating powdered food. At a body weight of 180-200 g, the animals were then randomly assigned to groups of 8-10 animals. The animals were housed in pairs in all metal cages. Throughout the "Experimental Period" of 6 days, the animals were fed ad-lib one of the following powdered diets: (a) Plain Purina Laboratory Chow -- (controls); or (b) Purina Chow containing THFES(HM). After the 6 days experimental period, the animals were exsanguinated by severing the external jugular veins under hexabarbital sodium acetate anesthesia (100 mg/kg I.P.) the blood being collected in acid-washed tubes to prevent alkaline hydrolysis of the triglycerides. The serum was separated by centrifugation, and the total cholesterol levels were estimated on the Auto-Analyzer using Standard Method No. 24a for cholesterol (Technicon Auto Analyses Procedures Manual). Body weight increases were computed from measurements of body weight at the beginning and at the end of the experimental period. Livers were dissected and weighed wet. Absolute liver weight and liver weight as a percentage of final body weight were recorded. Food consumption was monitored throughout the experimental period. The actual amount of drug ingested (in mg./kg./day) was computed from the food consumption, final body weight, and the known amount of drug in diet. Table I below shows the effects of THFES(HM) on the serum total cholesterol levels of the animals. Significant depression of serum total cholesterol was seen at doses of 6.9 mg./kg./day and above. THFES(HM) did not produce hepatomegaly under these conditions. F.E.S., THFES(LM), deoxy F.E.S. and p-methyl F.E.S. can be substituted for the THFES(HM) to treat cholesterol in the manner shown.

                  TABLE I                                                          ______________________________________                                                       Total Serum Cholesterol                                          Treatment Dose*     Mean     (mg.%) Change as %                                          (Mg/kg/day)                                                                              ±SEM         of Control                                 Control**           58.8 ±                                                                               1.7    --                                         ______________________________________                                         THFES (HM)                                                                               1.0       58.5 ±                                                                               3.3    +1.7                                       "         2.1       57.7 ±                                                                               2.4    0                                          "         6.9       49.5 ±                                                                               2.1    -13.8                                      "         8.5       46.6 ±                                                                               2.6    -19.0                                      "         13.2      41.6 ±                                                                               1.4    -27.6                                      "         16.4      38.1 ±                                                                               1.7    -34.5                                      "         30.0      33.5 ±                                                                               2.1    -41.4                                      "         53.0      33.5 ±                                                                               4.4    -41.4                                      "         100.0     24.5 ±                                                                               1.8    -57.0                                      ______________________________________                                          *Calculated from dietary concentration of drug and food consumption.           **Mean for two experiments.                                              

EXAMPLE XXV

Groups of 35-day old male rats (Holtzman) weighing 120-135 gm were held without treatment for 1 week on standard laboratory ration (Purina Labena Chow). The rats were then fed, ad libitum, for a 2-week period, a diet consisting of the following:

    Purina Labena     50.4%                                                        Cornstarch Mixture*                                                                              29.0                                                         Lard              20.0                                                         DL-Methionine     0.6                                                          *Cornstarch Mixture:                                                           Cornstarch         70%                                                         Alphacel          12                                                           Crisco            10                                                           USP Salt Mixture XIV                                                                              7                                                           Cod Liver Oil      1                                                      

During the next 2 weeks (4th and 5th on experimental diet) the animals received test compounds that were mixed in the above on a mg./kg. of diet basis. After exposure to this diet containing drug for 2 weeks the animals were anesthetized, bled individually to obtain serum samples to determine total cholesterol, triglycerides and α- and β- lipoproteins.

The results are set forth in Table II. The chemical analysis of sera of rats on the high fat diet for 4 weeks, with drugs added for the last 2 weeks of the test period, disclosed that THFES(HM) produced highly significant decreases in total cholesterol at all doses tested and THFES(HM) decreased the α/β lipoprotein ratio to highly significant degrees at the four concentrations tested; the α/β ratios decreased with increasing concentration in the diet.

While THFES(HM) produced marked decreases in the cholesterol, triglycerides and α/β- lipoprotein ratios, treatment with the compound also produced decreases in the weight gain of the animals. Based on the daily food intake values,

                                      TABLE II                                     __________________________________________________________________________     Compound                                                                              Dose                                                                               No.                                                                               Av. Food                                                                             Total         Lipo-        Testes                                 mg/kg                                                                              Rats                                                                              Intake                                                                               Cholesterol                                                                           Triglycerides                                                                         proteins                                                                              a/β                                                                             Wt. Body Weight*                       Diet   gm/day/rat                                                                           (mg/100 ml serum)                                                                            a-  β                                                                            Ratio (gm)                                                                               Init.                                                                              Start                                                                              Final               __________________________________________________________________________     Control                                                                               --  10 13    98.7 ± 4.0                                                                         321.8 ± 46.0                                                                        18.3                                                                              27.7                                                                              0.66  3.3 126 269 342                 THFES (HM)                                                                            12.5                                                                               10 14.7  72.7 ± 4.0***                                                                       214.2 ± 22.2*                                                                      15.8                                                                              32.5                                                                              0.49**                                                                               3.3 127 260 287                 THFES (HM)                                                                            25.0                                                                               10 13.5  62.5 ± 3.0***                                                                      258.1 ± 41.1                                                                        11.6                                                                              29.7                                                                              0.39**                                                                               3.2 128 264 278                 "      50.0                                                                               10 12.4  59.5 ± 5.4***                                                                      279.2 ± 40.2                                                                         7.2                                                                              32.8                                                                              0.22***                                                                              3.1 127 255 255                 "      100.0                                                                              10 11.0  52.0 ± 2.8***                                                                      185.4 ± 29.0                                                                         3.0                                                                              33.7                                                                              0.09***                                                                              3.1 128 260 246                 __________________________________________________________________________      *P <0.05                                                                       **P <0.01?                                                                     ***P <0.001                                                                     *Initial = Weight on day animals were received                                 Start = Weight on day animals were placed on diet plus compound.        

the estimated daily dose of THFES(HM) in these animals varied from 180 to 1100 g per rat. In addition to the findings discussed above, four of ten animals on the 12.5 mg dose level showed signs of alopecia; 1/10 of the high dose group presented this symptom. As has been indicated in other studies there was involution of the thymus in treated animals. F.E.S. and deoxy THFES can be substituted for the THFES(HM) to treat hypercholesterolemia in a similar manner.

The following examples demonstrate that the active ingredient of this invention is effective in estrogenic replacement therapy in post-menapausal and similar treatments.

EXAMPLE XXVI

The effect of THFES(HM) on the vaginal and uterine mucosa and sex skin changes of ovariectomized Rhesus monkeys was determined to demonstrate the post-menapausal effects of the drug. Monkeys were studied for a period of several days prior to drug treatment to establish base-line vaginal values. THFES(HM) was administered orally for 10 days. Observations on vaginal smear changes, coloration of the sex skin and withdrawal bleeding were recorded during treatment and for 20 days following its cessation.

Administration of the drug resulted in increased numbers of cornified cells in the vaginal washings. By the 5th day of treatment with 1.8 mg/kg, leukocytes were absent from the smear and remained absent for the duration of the treatment period. Coloration of the sex skin at this dose reached its maximum on the 4th day of treatment in animal No. 13 and the 11th day in animals No. 12 and No. 14. It would appear that, at this dose level, vaginal cornification is the most sensitive indicator of the estrogenic effect of the drug. The effect on the uterus is indicated by the withdrawal bleeding which occurred in two of three animals treated; there appeared to be little uniformity in the time at which bleeding occurred following cessation of treatment.

Four of six animals treated with 0.9 mg./kg. drug had withdrawal bleeding following cessation of treatment indicating uterine stimulation; "spotting" was noted in 5 of the 6 animals. This response would suggest that 0.9 mg./kg. dose was stimulating endometrial development but was inadequate to maintain it. Vaginal changes were comparable with those observed in animals treated with the 1.8 mg./kg. dose level. Sex skin changes were present and the degree of coloration was essentially the same as was observed in the high dose group.

Animals treated with 0.45 mg./kg. drug displayed minimal or no estrogen-dependent changes. Animals No. 28, No. 29 and No. 30 showed a vaginal response which persisted throughout the study. Animal No. 29 had "spotting" as judged by the presence of RBC's in its smear on 11 treatment or post-treatment days. Animals No. 23, No. 24 and No. 26 treated at a different time, failed to respond.

Based upon the data, it can be concluded that THFES(HM) is able to stimulate estrogen-like changes in ovariectomized monkeys and can be used as a post-menapausal drug.

EXAMPLES XXVII-XXX

In each of these examples, 10 women were administered THFES(HM) at varying dosage levels for a period of time. Tables III to VI set forth the relevant data for post-menapausal response. The administration of the drug and dosage was

    TABLE      DOSAGE         TEST PERIOD                                          ______________________________________                                         Table III      400 mg./daily  20 days                                          Table IV       200 mg./daily  20 days                                          Table V        100 mg./daily  60 days                                          Table VI        50 mg./daily  30 days                                          ______________________________________                                    

The prior symptoms appear with the patient identification and the effect of the drug is shown under Remarks. The data demonstrate the effectiveness of THFES(HM) in relieving many of the symptoms at the dosage levels tested. At 200 mg./daily the symptoms, with the exception of insomnia, were relieved. At 100 mg./daily the symptoms were relieved and the patients showed excellent estrogenic response with the exception of Patient 63-A. There were no toxic symptoms evident. With an average weight per patient of about 50 kg., the dosages levels are about 1 to 8 mg./day/kg. body weight. The follicle stimulating hormones (FSH) data of Table V are a clear demonstration of the activity of the drug in treatment of post-menapausal syndrome since to lower the FSH units in a good measure of such activity.

                                      TABLE III                                    __________________________________________________________________________                    Vaginal Cytology                                                                     Basal      Leuco-                                         Patient        Age                                                                               Day                                                                               Cells                                                                              CPI EI cytes    Remarks                               __________________________________________________________________________        39-A                                                                        Had vulvular pruritis                                                                         62 0  77% 23     Many Shortly after start - patient             and insomnia.     7  0   38  64 Some feeling well with slight de-              Diabetes mellitus in                                                                             14 0   56  61 v.few                                                                               crease of vulvar itching and              good control with 21 0       87 None wetness of vulva and vagina.              diet. Biopsy -    21 Biopsy - Late prolif.                                                                          Did not have withdrawal bleed-            resting endom.    endometrium. Moderate                                                                             ing. Discontinued - lack of                                 mitotic activity.  drug.                                        40-A                                                                        Suffers hot flushes                                                                           51 0  17   6  19 Some Complete disappearance of hot             and irritability. 7  0   32  56 Some flushes. Improvement of                   Biopsy - Early prolif,                                                                           14     45  39 v.few                                                                               psychological state. Dis-                 endom. Scarce mitotic                                                                            20     67  48 v.few                                                                               continued - lack of drug.                 activity.         20 Biopsy - Late prolif                                                                           Withdrawal bleeding lasted                                  endometrium. Patchy                                                                               3 days.                                                     hyperplasia.                                                    41-A                                                                        Periodic diarrhea and                                                                         59 0  34   2  10      Colon condition unchanged.                dryness of genitalia.                                                                            7  6   21  36      Genitalia moist. Has had nor-             Irritability of colon                                                                            14     59  83      mal intercourse with husband.             since age 30. Biopsy-                                                                            20     48  71      Scant withdrawal bleeding                 Resting endometrium.                                                                             20 Biopsy - Late prolif.                                                                          lasting 2 days. Discontinued -                              endometrium. Good  lack of drug.                                               mitotic activity.                                               42-A                                                                        Depression, irritabil-                                                                        52 0  11  19   6 Some Hot flushes and irritability dis-         ity and mild hot  7      24  51 Few  appeared. Still depressed. No             flushes. Biopsy - 14     52  60 Few  withdrawal bleeding. Discontinued -       Intermed. prolif. 20     63  57 Few  lack of drug.                             endom. Scarce      20 Biopsy - Late prolif.                                    mitotic activity. endometrium. Good mitotic                                                      activity.                                                       43-A                                                                        Obese; has insomnia                                                                           59 0  31   7  22      No headaches. Insomnia did not            and frequent head-                                                                               7  2   32  26      improve. Slight withdrawal                aches. Biopsy -   14     51  64      bleeding that lasted 2 days.              atrophic endom. with                                                                             20     59  47      Discontinued - lack of drug.              areas of cystic pseudo-                                                                          20 Biopsy - Late prolif.                                     hyperplasia.      endom. with moderate                                                           mitotic activity.                                               44-A                                                                        Hot flushes and dys-                                                                          51 0  24  12   3      Hot flushes disappeared. Dys-             parennia. Cholecis-                                                                              7  0   38  63      pareunia minimal. No with-                tectomy in 1961.  14     67  100     drawal bleeding. Discon-                  Biopsy - Intermed.                                                                               20     77  91      tinued - lack of drug.                    prolif. endom.    20 Biopsy - Late prolif.                                     Scarce mitotic activity.                                                                         endometrium. Good mitotic                                                      activity.                                                        45-A                                                                       Marked dryness of                                                                             63 0  87 marked inflammatory -                                                                       Markedly relieved - epithelia             genitalia - discrete   exudate       of the genitalia are moist -              burning sensation.                                                                               7  14  38  22      burning sensation disappeared.            Has moderate hyper-                                                                              14 0   63  71 Several                                                                             No withdrawal blending. Dis-              tension. Biopsy - 20     68  90 Some continued - lack of drug.                 atrophic endometrium.                                                                            20 Biopsy - Late prolif.                                                       endometrium. Moderate                                                          mitotic activity.                                               46-A                                                                        Easy fatigability and                                                                         56 0  18  13  22 Several                                                                             Marked decrease of irritability;          irritability. Biopsy -                                                                           7      39  63 Some still gets tired easily;                  resting endometrium.                                                                             14     68  70 Some Withdrawal bleeding lasted 4                                20     73  69 Few  days and was moderate. Dis-                                 20 Biopsy - Late prolif.                                                                          continued - lack of drug.                                   endometrium. Patchy                                                            hyperplasia.                                                    47-A                                                                        No gynecological                                                                              59 0  43  11   2 Some No ill effects - had slight               complaints -      7      43  90 Few  mucorrhea lasting 2 days.                 chronic colitis.  14     51  67 Few  Withdrawal bleeding moderate              Biopsy - Inactive 20     73  82 No   lasting 2 days. Discon-                   endometrium       20 Biopsy - Late prolif.                                                                          tinued - lack of drug.                                      endom. Moderate                                                                mitotic activity.                                               48-A                                                                        Abdominal discomfort                                                                          52 0      23  30 Some Abdominal discomfort and hot              rare; marked hot  7      53  68 v.few                                                                               flushes disappeared. Remains              flushes and moderate                                                                             14     87  82 v.few                                                                               depressed. No withdrawal                  depression.       20     79  94 v.few                                                                               bleeding. Discontinued -                  Biopsy -          20 Biopsy - Late prolif.                                                                          lack of drug.                             Late prolif. endom.                                                                              endometrium. Good                                            Slight mitotic ac-                                                                               mitotic activity.                                            tivity.                                                                        __________________________________________________________________________

                                      TABLE IV                                     __________________________________________________________________________                    Vaginal Cytology                                                                     Basal      Leuco-                                         Patient        Age                                                                               Day                                                                               Cells                                                                              CPI EI cytes    Remarks                               __________________________________________________________________________        49-A                                                                        Discrete depression                                                                           55 0       8   2 Few  Feels same as before starting             and fainting spells.                                                                             7      28  19 Few  drug. No withdrawal bleed-                Biopsy - Inactive 14     24  36 Few  ing. Discontinued - lack                  endometrium.      20     31  22 Few  of drug.                                                    20 Biopsy - intermed.                                                          prolif. endom. - Moderate                                                      mitotic activity.                                               50-A                                                                        Minor complaints on                                                                           60 0  61         Several                                                                             Condition remained the same               GF tract; bouts of                                                                               7  32  21   7      during treatment, although                diarrhea. Emotionally                                                                            14     53  67 Some genitalia became wet.                     unstable.         20     47  83 Some Discontinued - lack of drug.              Biopsy - atrophic 20 Biopsy - intermed.                                        endometrium.      prolif. endom. Moderate                                                        mitotic activity.                                               51-A                                                                        Slight itching and                                                                            65 0  68   2   7 Many Itching and Leukorrhea dis-               leukorrhea; irrita-                                                                              7      63  70 Some appeared. Psychological                   bility for past 20                                                                               14     56  82 Few  state did not change. Mini-               years.            20     59  67 Few  mal withdrawal bleeding for               Biopsy - none.    20 Biopsy - Late prolif.                                                                          one day. Discontinued -                                     endom. Moderate mitotic                                                                           lack of drug.                                               activity.                                                       52-A                                                                        Suffers dyspareunia,                                                                          54 0  4   23  32 Some Increased wetness of vagina               headaches, insomnia.                                                                             7      38  40 Few  and decrease of dyspareunia.              Biopsy - intermed.                                                                               14     30  31 Some Headache and insomnia not                 prolif. endometrium -                                                                            20     47  38 Few  affected. No withdrawal                   few mitoses.      20 Biopsy - Late prolif.                                                                          bleeding. Discontinued -                                    endometrium. Moderate                                                                             lack of drug.                                               mitotic activity.                                               53-A                                                                        Dryness of vulva                                                                              65 0  86         Many Complete disappearance of com-            and vagina - slight                                                                              7      32  53 Many plaints. Regrets no more                  burning sensation 14     67  84 Some drug available. No with-                  worse at night    20     73  69 Some drawal bleeding. Discon-                  Biopsy - atrophic 20 Biopsy - Late prolif.                                                                          tinued - lack of drug.                    endometrium       endometrium. Slight                                                            mitotic activity.                                               54-A                                                                        Marked irritability -                                                                         53 0  22  12   4 Some No hot flushes during treat-              infrequent but very                                                                              7  6   22  19 Some ment. First days of treat-                hot flushes       14     48  36 Some ment moderate nausea. One                 Biopsy - atrophic 20     36  49 Some episode of epigastric burn-               endom. - cystic   20 Biopsy - Late prolif.                                                                          ing. Irritability did not                 pseudohyperplasia.                                                                               endometrium. Moderate                                                                             change. No withdrawal                                       mitotic activity.  bleeding. Discontinued -                                                       lack of drug.                                55-A                                                                        Hot flushes and                                                                               50 0      15  23 Several                                                                             No hot flushes; palpitations              palpitations.     7      42  38 Few  unchanged. Epigastric                     Biopsy - Intermed.                                                                               14     59  64 Few  burning first few days.                   prolif. endom.    20     53  71 Few  Moderate withdrawal bleeding              Scarce mitotic    20 Biopsy - Late prolif.                                                                          lasted 3 days. Discontinued -             activity          endometrium. Slight                                                                               lack of drug.                                               hyperplasia.                                                    56-A                                                                        Dryness of vulva                                                                              58 0  73         Many Marked improvement with dis-              and marked dys-   7  22  19  24 Several                                                                             appearance of dyspareunia.                pareunia last 2   14     38  57 Some No withdrawal bleeding.                   years.            20     71  64 Few  Discontinued - lack of drug.              Biopsy - resting  20 Biopsy - Late prolif.                                     endometrium.      endometrium. Moderate                                                          mitotic activity.                                               57-A                                                                        Extremely irritable                                                                           53 0      10   0 Few  Hot flushes gone and feels                with frequent hot 7      48  30 Few  much better. Regrets no                   flushes           14     36  63 Few  more drug. Withdrawal                     Biopsy - Intermed.                                                                               20     57  75 Few  bleeding moderate - lasted                prolif. endom.    20 Biopsy - Late prolif.                                                                          two days. Discontinued -                  Very few mitoses. endometrium. Good  lack of drug.                                               mitotic activity.                                               58-A                                                                        Palpitation, moder-                                                                           64 0  74%  3   0 Many Patient remained in same state.           ate respiratory in-                                                                              7  16  28  12 Several                                                                             No withdrawal bleeding. Dis-              sufficiency, easy 14     31  42 Several                                                                             continued - lack of drug.                 fatigability. Widow.                                                                             20     57  49 Some                                           Biopsy - none     20 Biopsy - Late prolif.                                                       endometrium. Slight                                                            mitotic activity.                                            __________________________________________________________________________

                                      TABLE V                                      __________________________________________________________________________     Vaginal Cytology                                                                                                  Follicle                                                       Basal      Leuco-                                                                              Stimulating                                 Patient      Age                                                                               Day                                                                               Cells                                                                              CPI EI cytes                                                                               Hormone Remarks                             __________________________________________________________________________       59-A                                                                                      52 0  64% 6      Several                                                                             0d. 52 m.u.                                                                            Under treatment 50 days,            Hot blushes; nausea                                                                            10 11  27  47 Some 45d. 6 m.u.                                                                            after 10 days symptoms              in the morning. 20     63  54 Some         disappeared. No prob-               Biopsy - resting                                                                               30     50  62 Some         lems.                               endometrium.    30 Biopsy - Prolif. endom.                                                     Moderate mitotic activity.                                       60-A                                                                                      50 0      7   16 Several                                                                             0d. 112 m.u.                                                                           Under treatment 54 days.            Asthenia and numbness                                                                          10     35  49 Several                                                                             60d. 12 m.u.                                                                           Marked improvement 10               of hands. Menopause                                                                            20     59  37 Several      to 15 days after start.             4 years ago. Biopsy -                                                                          30     68  49 Some         No problems.                        Early prolif. endom.                                                                           30 Biopsy - Late prolif.                                       Scarce mitotic  endometrium. Moderate                                          activity        mitotic activity.                                                61-A                                                                                      49 0      5   0  Many         Symptoms disappeared after          Hot flushes and slight                                                                         10     27  55 Several      2 weeks. No problems.               irritability. Biopsy -                                                                         20     63  42 Several                                          Resting endometrium.                                                                           30     58  60                                                                  30 Biopsy - Late prolif.                                                       endometrium. Slight                                                            mitotic activity.                                                62-A                                                                                      51 0  87% 3      Many 0d. 64 m.u.                                                                            Symptoms improved 6 or 7            Asthenia and irrita-                                                                           10 2   19  24 Some 30d.  6 m.u.                                                                           days after start. As-               bility. Biopsy -                                                                               20     63  40 Some 60d. >6 m.u.                                                                           thenia gone after 15                None.           30     51  66 Some         days. Irritability                                  30 Biopsy - Late prolif.   markedly decreased. No                              endometrium. Moderate      problems.                                           mitotic activity.                                                63-A                                                                                      57 0  93% 0      Many 0d. 120 m.u.                                                                           Hot flushes disappeared             Hot flushes and 10 22  30  12 Several                                                                             30d. 10 m.u.                                                                           after 7 to 10 days.                 itching of genitalia.                                                                          20     36  43 Several      Itching decreased but               Biopsy - resting                                                                               30     42  35 Several      did not vanish. After               endometrium.    30 Biopsy - Intermed. prolif.                                                                             2 weeks tenderness of                               endometrium. Slight        breasts and bloating.                               mitotic activity.          She interrupted treat-                                                         ment.                                 64-A                                                                                      55 0      9   16 Several                                                                             0d. 97 m.u.                                                                            Hot blushes gone in less            Hot blushes and 10     35  55 Several                                                                             30d. 9 m.u.                                                                            than 7 days. Three                  sporadic diarrhea.                                                                             20     45  38 Several                                                                             60d. 6 m.u.                                                                            of diarrhea. No                     Biopsy - Early prolif.                                                                         30     56  50 Several      other problem.                      endom. Minimal  30 Biopsy - Late prolif.                                       mitotic activity.                                                                              endometrium. Good mitotic                                                      activity.                                                        65-A                                                                                      63 0  78% 4      Several                                                                             0d. 185 m.u.                                                                           Hot blushes disappeared             Hot blushes and 10 32  16  10 Several                                                                             30d. <6 m.u.                                                                           and insomnia improved.              insomnia. Biopsy -                                                                             20 3   32  38 Some 60d. <6 m.u.                                                                           Feeling very well.                  resting endometrium.                                                                           30     29  62 Some                                                             30 Biopsy - Intermed. endom.                                                   Moderate mitotic activity.                                       66-A                                                                                      64 0  86% 0      Many 0d. 97 m.u.                                                                            Decrease in number of               Moderate irritability                                                                          10 33  30  17 Many 60d. 12 m.u.                                                                           headaches and intensity.            and headaches, also                                                                            20     33  46 Several      Mild nausea first                   mild diabetes mellitus.                                                                        30 6   45  32 Several      days. Vomited once.                 Biopsy - None.  30 Biopsy - Late prolif.                                                       endometrium. Slight                                                            mitotic activity.                                                67-A                                                                                      49 0      10  2  Some 0d. 106 m.u.                                                                           Hot blushes and head-               Hot blushes, nervous-                                                                          10     28  45 Some 7d. 31 m.u.                                                                            aches disappeared                   ness, sporadic head-                                                                           20     62  57 Few  14d. 25 m.u.                                                                           shortly after start.                aches. Menopause -                                                                             30     58  83 Few  21d. 13 m.u.                                                                           Nervousness improved                2 years ago. Biopsy -                                                                          30 Biopsy - Late prolif.                                                                          28d. 6 m.u.                                                                            but not gone.                       Late prolif. endom.                                                                            endometrium. Patchy                                                                               60d. <6 m.u.                                Scarce mitotic  hyperplasia.                                                   activity.                                                                        68-A                                                                                      34 0  98%        Several                                                                             0d. 168 m.u.                                                                           Feeling well. Symptoms              Complete hysterectomy                                                                          10 13  37  52 Some 30d. 14 m.u.                                                                           gone after 1.0 week.                and ovariectomy.                                                                               20 6   50  43 Some 60d. <6 m.u.                                Had severe      30     67  80 Few                                              hot blushes and 30 Biopsy - None                                               irritability. Biopsy-                                                          not done (hysterectomy)                                                        __________________________________________________________________________

                                      TABLE VI                                     __________________________________________________________________________     Vaginal Cytology                                                                                  Basal      Leuco-                                           Patient      Age                                                                               Day                                                                               Cells                                                                              CPI EI cytes     Remarks                                __________________________________________________________________________       79-A                                                                                      52 0  42% 6      Several                                                                             Hot blushes gone after 3 weeks.             Hot blushes and 10 23  14  28 Several                                                                             Remains very nervous.                       nervousness.    20     26  20 Several                                          Biopsy - atrophic                                                                              30     46  35 Several                                          endometrium. Cystic                                                                            30 Biopsy - Intermed. prolif. -pseudohyperplasia. endom.                       Moderate mitotic                                                               activity.                                                        80-A                                                                                      64 0  93%        Many Patient feels same and reports              Diarrhea (chronic                                                                              10 37  11     Several                                                                             no change.                                  amoebiasis).    20 30  28     Several                                          Depressed.      30 0   37  28 Several                                          Biopsy - resting                                                                               30 Biopsy - Early prolif.                                      endometrium.    endometrium. Moderate                                                          mitotic activity.                                                81-A                                                                                      50 0      11  31 Several                                                                             Hot blushes gone after 15-20                Marked irritability                                                                            10     35  30 Several                                                                             days. No headaches and moder-               frequent hot    20     63  48 Some ate decrease in irritability.               flushes, some severe                                                                           30     57  62 Some                                             headaches.      30 Biopsy - Late prolif.                                       Biopsy - resting                                                                               endometrium. Good mitotic                                      endometrium.    activity.                                                        82-A                                                                                      56 0  69% 0      Many Patient has not improved.                   Irritability,   10 31  27     Several                                          mild hot blushes and                                                                           20 8   19     Several                                          insomnia - past 2 years.                                                                       30     38  51 Some                                             Diabetic.       30 Biopsy - Late prolif.                                       Biopsy - resting                                                                               endometrium. Patchy                                            endometrium     hyperplasia.                                                     83-A                                                                                      69 0  100%                                                                               0      Many Good condition. Vaginal in-                 Atrophic vaginitis.                                                                            10 71  20     Many flammation improved.                        Hysterectomy in 20 17  48     Several                                          1953 Marked vari-                                                                              30 4   37     Several                                          cose veins.     30 Biopsy - None                                               Biopsy - hysterectomy                                                            84-A                                                                                      56 0      18  7  Several                                                                             Condition same as before start              Periods of mild 10     36  38 Some of treatment.                               depression or   20     31  54 Few                                              irritability.   30     40  36 Few                                              Biopsy - Early  30 Biopsy - Late prolif.                                       prolif. endom.  endometrium. Moderate                                          Scarce mitotic  mitotic activity.                                              activity.                                                                        85-A                                                                                      52 0  21% 13  6  Several                                                                             Slight improvement of all                   Intense, frequent                                                                              10     39  16 Some symptoms 10- 15 days after                  hot blushes. Occa-                                                                             20     36  44 Some start of drug.                              sional nausea and                                                                              30     42  29 Some                                             mild headaches. 30 Biopsy - Late prolif.                                       Biopsy - Not    endometrium. Patchy                                            enough material.                                                                               hyperplasia.                                                     86-A                                                                                      53 0      12  3  Several                                                                             Hot blushes disappeared. In-                Insomnia, moderate                                                                             10     36  51 Some somnia and nervousness un-                  depression, mild                                                                               20     27  50 Some changed.                                    occasional hot  30     47  26 Some                                             blushes. Biopsy -                                                                              30 Biopsy - Late prolif.                                       Early prolif. endom.                                                                           endometrium. Moderate                                          Scarce mitotic activity.                                                                       mitotic activity.                                                87-A                                                                                      66 0  96% 0      Many Vaginitis improved but has not              Kraurosis vulvae,                                                                              10 70  16     Many disappeared.                                atrophic vaginitis                                                                             20     35  52 Many                                             with moderate   30     28  27 Many                                             leukorrhea.     3 Biopsy - Intermed.                                           Biopsy - mocoid prolif. endom. Slight                                          material.       mitotic activity.                                                88-A                                                                                      49 0      11  21 Some Hot blushes gone 3 weeks after              Marked frequent hot                                                                            10     30  55 Few  start. Nervousness the same.                blushes and moderate                                                                           20     33  38 Few                                              nervousness.    30     28  47 Few                                              Biopsy - Early prolif.                                                                         30 Biopsy - Late prolif.                                       endom. Slight mitotic                                                                          endometrium. Poor                                              activity        mitotic activity.                                              __________________________________________________________________________

The terms "hot blushes" and "hot flushes" are used synonymously herein.

EXAMPLES XXXI and XXXII

Tables VI and VII below set forth the results of tests in which two groups of 10 women were fed, respectively, 200 mg./day and 100 mg./day of F.E.S. In the test summarized in Table VI the drug was administered daily for 60 days whereas in the test summarized in Table VII the drug was administered in a regime of 25 days on the drug followed by a 10 day rest period over a total time period of 180 days. Treatment at 100 mg./daily relieved most of the post-menapausal symptoms with the exception of insomnia and depression. Treatment at 200 mg./daily did not relieve insomnia.

                                      TABLE VII                                    __________________________________________________________________________     Vaginal Cytology                                                                                    Basal      Leuco-                                         Patient        Age                                                                               Day                                                                               Cells                                                                              CPI EI cytes     Remarks                              __________________________________________________________________________        139-A                                                                                      61 0  99% 0   0  Many Disappearance of symptoms re-             Suffers mild, well-                                                                              10 62  20  7  Many lated to genitalia. Head-                 controlled diabetes.                                                                             20 31  27  19 Several                                                                             aches unaffected. No                      Has headache and dry-                                                                            30 7   30  19 Several                                                                             withdrawal bleeding.                      ness with itching of                                                                             60 Late prolif. endom.                                       external genitalia.                                                                              Good mitotic activity.                                          140-A                                                                                      39 0  100 0   0  Many Patient well. No mor hot                  Had ovariohysterec-                                                                              10 52  26  13 Some blushes and feels very re-                tomy 8-67. Early  20 21  38  54 Some laxed. No withdrawal                      in October had hot                                                                               30 3   42  80 Few  bleeding.                                 blushes and nervousness.                                                          141-A          Early prolif. endom.                                                           Scarce mitotic activity.                                                    56 0      3   11 Several                                                                              Feels better but still has in-           Suffers insomnia and                                                                             10     26  31 Several                                                                             somnia. After second course               slight depression.                                                                               20     38  63 Some had few drops of withdrawal                                 30     46  45 Some bleeding.                                                   Late prolif. endom.                                                            Moderate mitotic activity.                                      142-A                                                                                         Resting endom.                                                              56 0  26  4   0  Several                                                                             No hot blushes since treatment            Marked nervousness                                                                               10 13  16  36 Some started. Nervousness im-                  and occasional hot                                                                               20 2   37  29 Some proved moderately.                        blushes.          30 0   46  38 Some                                                             Intermed. prolif. endom.                                                       Discrete mitotic activity.                                      143-A                                                                                      57    Atrophic endom. with                                                           pseudohyperplasia.                                                          0  17  2   9  Few  No change. No withdrawal                  Mild nervousness. 10     26  33 Few  bleeding.                                                   20     29  57 Few                                                              30     40  58 Few                                                              30 Intermed. prolif. endom.                                                       Moderate mitotic activity.                                   144-A                                                                                      69    No biopsy.                                                Suffered pyelonephritis                                                                          0  100 0   0  Several                                                                             Patient has mild renal insuffi-           and has atrophy and                                                                              10 64  10  7  Several                                                                             ency. Genitalia wetter with               dryness of vulva. 20 30  29  18 Several                                                                             occasional slight mucorrhea.                                30 7   35  52 Several                                                                             No withdrawal bleeding.                                     60 Early prolif. endom.                                                           Slight mitotic activity.                                     145-A                                                                                            Early prolif. endom.                                                           Discrete mitotic activity.                                Last menstrual period                                                                         49 0      13  26 Some No more hot blushes after one             (LMP) 5/14. Has frequent                                                                         10     35  30 Some week on drug. After end of                hot blushes and feels                                                                            20     48  39 Few  second course had vaginal                 nervous. On drug 11/14                                                                           30     66  50 Few  bleeding which was discrete.                                60 Late prolif. endom.                                                            Patchy hyperplasia.                                          146-A                                                                                            Early prolif. endom.                                                           Slight mitotic activity                                   Moderate nervousness                                                                          54 0  14  6   23 Several                                                                             Feels better but has had two              and bouts of diarrhea                                                                            10     21  29 Some episodes of diarrhea. After                                 20     38  34 Some second course had moderate                                  30     35  62 Some withdrawal bleeding for two                                                    days.                                        147-A             Resting endometrium.                                                     51 0  66  3   0  Several                                                                             Hot blushes disappeared shortly           LMP 13 mo. prior to drug.                                                                        10 9   21  38 Some after started drug and irri-              Has frequent hot blushes                                                                         20 7   39  32 Some tability improved slightly.               and marked irritability.                                                                         30 1   46  52 Some No withdrawal bleeding.                                     60 Intermed. prolif. endom.                                                       Slight mitotic activity.                                     148-A                                                                                            No biopsy.                                                Has mild diabetes                                                                             65 0  100 0   0  Many +                                                                              Besides drug patient received             and atrophic vulvular           oandida                                                                             anti-monilial drugs. Vulvo-               vaginitis.                      albicans                                                                            vaginitis cured. No with-                                   10 54  3   17 Many drawal bleeding.                                            20 20  35  23 Several                                                          30 13  56  60 Several                                        __________________________________________________________________________

                                      TABLE VIII                                   __________________________________________________________________________     Vaginal Cytology                                                                                   Basal     Leuco-                                           Patient       Age                                                                               Day                                                                               Cells                                                                              CPI                                                                               EI cytes     Remarks                                __________________________________________________________________________       119-A                                                                                      62 0  79% 0  2  Many Itching improved after one week and         Has hypertension and                                                                            10 36  7  10 Several                                                                             disappeared after 3 weeks. No               intense vulvar itching.                                                                         20 17  30 21 Several                                                                             withdrawal bleeding but some                Has diabetes mellitus                                                                           30 6   37 47 Some mucorrhea. Schedule: 25 days                controlled by dieting.                                                                          Endometrial biopsy after 20                                                                      on drug - 10 days off. 180 day              No endometrial biopsy                                                                           days - Intermediate prolif.                                                                      Biopsy - Late Proliferative en-             before treatment.                                                                               endometrium. Slight                                                                              dometrium. Good mitotic activity.                            activity.         At 180 days - Patient feeling                                                  well. Slight withdrawal bleeding                                               for 3 days after four months of                                                treatment.                                    120-A                                                                                      53 0  9   10 3  Some Marked improvement after 5 to 10 days       Infrequent hot blushes                                                                          10     23 36 Some of treatment. She has been                  and moderate nervousness.                                                                       20     51 67 Few  feeling well. Moderate with-                Biopsy - intermed. prolif.                                                                      30     40 58 Few  drawal bleeding after second course         endom. with slight                                                                              20 day Biopsy - Late pro-                                                                        of treatment. Schedule: 25 days             mitotic activity.                                                                               lif. endom. with moderate                                                                        on drug - 10 days off. At 180                                mitotic activity. days - Biopsy - Late proliferative                                             endometrium with good mitotic                                                  activity. Feeling well. From                                                   third to sixth cycle no withdrawal                                             bleeding.                                     121-A                                                                                      58 0  14  3  13 Several                                                                             Headache improved but not insomnia.         Has frequent headaches                                                                          10 3   26 17 Some Few drops of blood after third              and insomnia.    20     38 62 Few  course. Schedule: 25 days on                Biopsy - atrophic                                                                               30     31 70 Few  drug - 10 days off. Biopsy -                endom. with cystic                                                                              60 day Biopsy - prolif.                                                                          180 days - Proliferative en-                pseudohyperplasia.                                                                              endom. with patchy hyper-                                                                        dometrium with patchy hyperplasia.                           plasia.           180 days - Slight withdrawal                                                   bleeding on fourth and sixth                                                   cycles. Feeling well.                         122-A                                                                                      61 0  64  1  13 Many Feeling a little better. No with-           Moderate irrita- 10 24  27 42 Few  drawal bleeding. Schedule: 25               bility and depression.                                                                          20 18  18 58 Few  days on drug - 10 days off. 180             Biopsy - inactive                                                                               30 3   40 37 Few  days biopsy - Late proliferative            endometrium.     30 day Biopsy - prolif.                                                                          endometrium. Feels depressed-                                endom. Slight mitotic                                                                            no withdrawal bleeding.                                      activity.                                                       123-A                                                                                      50 0      16 24 Some After one week of treatment, hot            Frequent intense hot                                                                            10     46 37 Some blushes decreased in number                 blushes with abdominal                                                                          20     35 58 Few  and intensity and disappeared               pain and flatulence.                                                                            30     58 65 Few  after 20 days. Flatulence has               Biopsy - early prolif.                                                                          60 day biopsy - Late pro-                                                                        decreased but not disappeared.              endom. with poor lif. endom. with good                                                                            Schedule: 25 days on drug - 10              mitotic activity.                                                                               mitotic activity. days off. 180 day biopsy - Late                                                -  Proliferative endometrium with                                              moderate mitotic activity.                                                     Patient has episodes of abdominal                                              discomfort, otherwise feels well.                                              No withdrawal bleeding.                       124-A                                                                                      68 0  98  0  0  Many Itching improved; vulvar mucosa             Kraurosis vulvae and                                                                            10 80  0  7  Many slightly wet. No withdrawal                 moderate itching.                                                                               20 53  11 28 Several                                                                             bleeding. Schedule: 25 days on              Biopsy - resting 30 21  35 31 Several                                                                             drug - 10 days off. 180 day                 endometrium.     30 day biopsy - Early pro-                                                                       biopsy - Intermediate prolifera-                             lif. endom, with slight                                                                          tive endometrium with poor mitotic                           mitotic activity. activity. Feeling well. No with-                                               drawal bleeding.                              125 A                                                                                      52 0      8  3  Some No headaches and hot blushes dis-           Moderately frequent hot                                                                         10     51 34 Some appeared after one week on                  blushes; rare headaches                                                                         20     35 62 Some drug. Minimal withdrawal bleeding           Biopsy - atrophic endom.                                                                        30     60 48 Some after third course. Schedule:               with cystic pseudo-                                                                             30 day biopsy - prolif.                                                                          25 days on drug - 10 days off.              hyperplasia.     endom. with moderate                                                                             180 day biopsy - Late prolifera-                             mitotic activity. tive endometrium with patchy                                                   hyperplasia. Occasional head-                                                  aches fifth month and slight                                                   withdrawal bleeding on fifth                                                   month.                                        126-A                                                                                      58 0  47  1  13 Many Irritability improved but still de-         Irritability and periods                                                                        10 37  8  20 Several                                                                             pressed over family problems.               of depression.   20 26  17 19 Several                                                                             No withdrawal bleeding. Schedule:           Biopsy - resting en-                                                                            30 8   31 43 Several                                                                             25 days on drug - 10 days off.              dometrium.       30 day biopsy - Early pro-                                                                       180 day biopsy - Late prolifera-                             lif. endom. Moderate mito-                                                                       tive endometrium with discrete                               tic activity.     mitotic activity. Depressed. Has                                               forgotten to take several capsules.                                            No withdrawal bleeding.                       127-A                                                                                      55 0  37  8  3  Several                                                                             Remained without significant                                 10 20  17 29 Some change; likely nervousness                  Slight nervousness                                                                              20 7   35 23 Some decreased. No withdrawal                    and hypertension.                                                                               30 11  47 61 Few  bleeding. Schedule: 25 days                 Biopsy - resting 60 day biopsy - late pro-                                                                        on drug - 10 days off. 180                  endometrium.     liferative endometrium.                                                                          day biopsy - Early prolifera-                                Moderate mitotic activity.                                                                       tive endometrium with moderate                                                 mitotic activity. Irritability                                                 improved. No withdrawal                                                        bleeding.                                     128-A                                                                                      49 0      14 26 Many Symptoms improved in five days and          Marked and frequent                                                                             10     23 31 Many disappeared after 15 days. Feel-            hot blushes with fre-                                                                           20     28 63 Several                                                                             ing well. Moderate withdrawal               quent moderate headaches.                                                                       30     55 42 Several                                                                             bleeding after second and third             Biopsy - early prolif.                                                                          30 day biopsy - late prolif.                                                                     courses. Schedule: 25 days on               endom. with moderate                                                                            endometrium. Good mitotic                                                                        drug - 10 days off. 180 day                 mitotic activity.                                                                               activity.         biopsy - Proliferative endometrium                                             with patchy hyperplasia. Feeling                                               well. Moderate withdrawal bleeding                                             in fifth and sixth months.                  __________________________________________________________________________

EXAMPLE XXIII

Groups I, II, III and IV each composed of human female patients suffering postmenapausal syndrome were orally given dosages of 10, 25, 50 and 100 mgs./day, respectively, of pure Compound II (LM) having a melting point of about 155°C. for 3 cycles, each cycle consisting of taking the compound daily for 20 days followed by a 10 day period during which the patient abstains from taking the compound. The results are set forth below.

The pure Compound II (LM) can be separated from a mixture of it and its high melting diastereoisomer by solubilizing the mixture in glacial acetic acid and crystallizing out the pure Compound II (LM) in accordance with the process described in the copending patent application of Vernon V. Young, Ser. No. 643,819, filed June 6, 1967, herein incorporated by reference.

              FSH (units).sup.2   VAGINAL CYTOLOGY.sup.3                                                                        SUBJECTIVE                                                      CPI            RESPONSE.sup.4                    GROUP                                                                               DOSE.sup.1                                                                          INITIAL 90 DAYS TREATMENT                                                                          INITIAL                                                                               20-DAYS 90 Days                           __________________________________________________________________________     I    10 mg.                                                                              105(144-66)                                                                              25.8(54-6)                                                                               5.2(18-0)                                                                             24.7(38-16)                                                                            4 of 10                           II   25 mg.                                                                              102(156-66)                                                                              22.2(36-6)                                                                               7.6(19-0)                                                                             40.8(52-26)                                                                            8 of 10                           III  50 mg.                                                                              92.8(138-54)                                                                             12.0(30-6)                                                                               5.5(11-0)                                                                             55.2(73-42)                                                                            10 of 10                          IV   100 mg.                                                                              99(150-60)                                                                              11.4(30-6)                                                                               11.0(19-0)                                                                            67.5(77-59)                                                                            9 of 10                           __________________________________________________________________________      .sup.1 Dose mg. orally daily for 20 days with a 10 day rest.                   .sup.2 Follicle stimulating hormone assay before and after 3 cycles of         treatment.                                                                     .sup.3 Chorionic pycnotic index.                                               .sup.4 Subjective response as reported to physician by patient.           

It is claimed:
 1. A method of treating post-menapausal syndrome in a human female having such syndrome which comprises internally administering to said human an amount sufficient to relieve said syndrome of a compound of the formula selected from the group consisting of: ##SPC7##
 2. The method of claim 1 wherein the compound is orally administered.
 3. The method of claim 2 wherein the compound is administered in a cycle of 20 to 25 days of administration followed by a rest period of 5 to 10 days.
 4. The method of claim 3 wherein the compound is administered to said human in an amount of about 0.1 to 10 mg. per day per kilogram body weight of said human.
 5. The method of claim 4 wherein said compound is the diastereoisomer of the formula ##SPC8##having a melting point of about 155°C.
 6. The method of claim 1 wherein said compound is the diastereoisomer of the formula ##SPC9##having a melting point of about 178°-180°C. 